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題 名 | 應用反轉錄聚合酶連鎖反應鑑定暨區分新城雞病病毒疫苗株及野外株=Identification and Differentiation of Vaccine Strains and Field Isolates of Newcastle Disease Virus by Reverse Transcription-Polymerase Chain Reaction |
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作 者 | 張伯俊; 黃志銘; 楊程堯; 謝快樂; | 書刊名 | 中華民國獸醫學會雜誌 |
卷 期 | 23:6 1997.12[民86.12] |
頁 次 | 頁546-555 |
分類號 | 437.717 |
關鍵詞 | 新城雞病; 反轉錄聚合酶鏈反應; Newcastle disease; Reverse transcription-polymerase chain reaction; |
語 文 | 中文(Chinese) |
中文摘要 | 為了鑑定區分新城雞病病毒疫苗株與野外強毒株,我們利用反轉錄聚合�t鏈反應 技術,建立快速區分新城雞病病毒的方法。首先,我們選擇了一組對新城雞病病毒具專一性 的引子進行 RT-PCR, 當待測物含有病毒時,我們可增幅出一段 426 base pairs (bp) 產物 ,再將此產物分別以限制�t HinP1 I 與 HaeIII 作用,臺灣株只會被 HinP1 I 作用,而疫 苗株則只能被 HaeIII 作用,藉此可區分新城雞病病毒疫苗株與野外株。另一方面,我們又 設計了三段對新城雞病病毒具專一性的引子,進行多引子反轉錄聚合�t鏈反應,若病材含有 新城雞病病毒野外株時,就可增幅一段 724bp 產物, 反之,若有疫苗株存在時,則會增幅 出 349bp 的產物, 因此也可區分新城雞病病毒疫苗株與野外株。 由本實驗的結果證明, RT-PCR 配合限制�t切割, 或直接進行多引子 RT-PCR 反應,皆可應用於新城雞病病毒之快 速區別診斷。 這是第一篇應用多引子 RT-PCR 來診斷新城雞病病毒之報告。[* 張白俊、黃 志銘、楊程堯、謝快樂。應用反轉錄聚合�t連鎖反應鑑定暨區分新城雞病病毒疫苗株及野外 株。中華獸醫誌 23(6):546-555,1997。* 聯絡人 TEL:04-2860196,FAXL:04-2851741] |
英文摘要 | We developed a rapid and simple procedure, based on the reverse transcription-polymerase chain reaction (RT-PCR), to identify and differentiate Taiwanese field isolates and vaccine strains of Newcastle disease virus(NDV). A 426 base-pairs (bp) fragment was amplified from samples containing NDV by RT-PCR, and then digested by restriction endonucleases HinP1 I and Hae III. The fragment amplified from Taiwanese isolates could be digested by HinP1 I but not by HaeIII. In contrast, the fragment amplified from vaccine strains could be digested by Hae III but not by HinP1 I. Therefore, RT-PCR combined with restriction enzyme digestion provides a convenient procedure for discriminating Taiwanese field isolates from vaccine strains of NDV. In addition, we developed a multiple-primers RT-PCR procedure for the identification and differentiation of NDV. In this procedure, three primers were included in the RT-PCR, so that a 724 bp fragment of HN gene was amplified from samples containing Taiwanese isolates, while a 349 bp fragment was amplified from samples containing vaccine strains. In this way, we could differentiate field isolates of NDV from vaccine strains by a simple RT-PCR assay. Results of this study demonstrate that RT-PCR provides a simple and rapid method for the detection and differential diagnosis of NDV. This is the first report on using multiple-primers RT-PCR in the diagnosis of NDV. [*Chang PC, Hwang JM, Yang CY and Shieh Happy K. Identification and differentiation of vaccine strains and field isolates of Newcastle disease virus by reverse transcription-polymerase chain reaction. J Chin Soc Vet Sci 23(6):546-555,1997. *Corresponding author TEL:04-2860196, FAX:04-2851741] |
本系統中英文摘要資訊取自各篇刊載內容。